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TGF-β, essential for development and immunity, is expressed as a latent complex (L-TGF-β) non-covalently associated with its prodomain and presented on immune cell surfaces by covalent association with GARP. Binding to integrin αvβ8 activates L-TGF-β1/GARP. The dogma is that mature TGF-β must physically dissociate from L-TGF-β1 for signaling to occur. Our previous studies discovered that αvβ8-mediated TGF-β autocrine signaling can occur without TGF-β1 release from its latent form. Here, we show that mice engineered to express TGF-β1 that cannot release from L-TGF-β1 survive without early lethal tissue inflammation, unlike those with TGF-β1 deficiency. Combining cryogenic electron microscopy with cell-based assays, we reveal a dynamic allosteric mechanism of autocrine TGF-β1 signaling without release where αvβ8 binding redistributes the intrinsic flexibility of L-TGF-β1 to expose TGF-β1 to its receptors. Dynamic allostery explains the TGF-β3 latency/activation mechanism and why TGF-β3 functions distinctly from TGF-β1, suggesting that it broadly applies to other flexible cell surface receptor/ligand systems.

Background: Pulmonary fibrosis (PF) is a chronic interstitial lung disease posing significant health risks. This study aimed to investigate the therapeutic mechanism of Qingfei Tongluo mixture (QTm) in treating PF by combining network pharmacology and experimental verification. Results: A total of 246 active ingredients in QTm were identified, with 159 potential targets for PF treatment. Quercetin, a key active ingredient, was associated with the TGF-b1 signaling pathway. Gene Ontology and KEGG enrichment analyses identified 42 core genes, with a notable implication of the TGF-beta signaling pathway in PF. Immunohistochemistry showed elevated FTO and TGF-b1 levels in PF tissues. Animal experiments demonstrated that QTm improved alveolar structure, reduced interstitial lesions, and enhanced lung function while decreasing hydroxyproline content and the expression of FTO and TGF-b1 proteins. Conclusions: QTm may inhibit PF progression by suppressing FTO/TGF-b1 expression, thereby improving lung function. These findings suggest that QTm holds potential as a treatment for PF. [ABSTRACT FROM AUTHOR]

Background: The aim of the study was to examine the significance of plasma Transforming Growth Factor-1/TGF-b1 (TGF-b1) level testing in patients with Type 2 Diabetes Mellitus (T2DM) and heart failure. Methods: A sample of T2DM patients who were hospitalised for dyspnea was chosen between June 2021 and June 2023. Based on the convenience sample approach, 150 cases were screened for the study, and 50 healthy non-diabetic people without cardiac problems who completed physical examinations over the same period were included as a control group. All study participants had their serum NT-proBNP and plasma TGF-I levels checked, and the values between the two groups were compared. Then, the patients with T2DM with heart failure were grouped according to whether they were accompanied by heart failure or not and the grading of cardiac function, and then the serum NT-proBNP and plasma TGF-b1 levels were compared between the different groups of patients. The diagnostic value of plasma TGF-b1 in the occurrence of heart failure in patients with T2DM was analysed. Results: There were 54 patients without heart failure and 96 people with heart failure among the 150 T2DM patients. The cut-off point was 44.50 g/L. At this time, the sensitivity and specificity for diagnosing concomitant heart failure in T2DM were 79.63% and 52.51%, respectively. 96 individuals with T2DM and heart failure showed greater serum and plasma levels of NT-proBNP and TGF-b1 compared to the other two groups (P=0.05). ProBNP and plasma TGF-b1 levels had a positive and significant relationship (P=0.05). Conclusions: Plasma TGF-b1 levels were much higher in T2DM patients than in the general population, and the increase in this index was more pronounced in patients who also had heart failure, a diagnostic indicator for T2DM and heart failure.

The investigation of novel phytochemicals for the prevention and treatment of infections caused by multidrug-resistant pathogens is gaining attention. The current study evaluated the in-vitro antimicrobial activity of kaempferol against methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. Its in-vivo efficacy in inhibiting these pathogens was determined using an excision wound model in nicotinamide-streptozocin- -induced diabetic rats. Kaempferol displayed an inhibitory effect against the tested bacteria both in vitro and in vivo. It also healed excision wounds at a 1% (w/w) concentration. An increase in antioxidant enzymes in wounded tissue was observed on kaempferol treatment. A reduction in the MRSA and P. aeruginosa counts in wounded tissue together with a reduced epithelization period was observed when compared to the infected control. A thicker epithelium, new capillaries, and a decrease in inflammatory cells were detected by hematoxylin and eosin staining. Furthermore, an increase in collagen fibers and their deposition was observed by Masson’s trichrome staining. Kaempferol at 40 µM did not display any toxicity for human keratinocytes grown in media containing high glucose and it did not affect the expression of the pro-healing cytokines genes vascular endothelial growth factor (VEGF) and transforming growth factor-b-1 (TGFβ1). Kaempferol displayed antibacterial and antioxidant actions but did not increase the expression of proliferative genes.

Despite Non-small cell lung cancer (NSCLC) ranks among the most deadly cancers worldwide, and currently, apart from a low percentage, targetable molecules have not been identified in its etiopathogenesis. The relationship between the proteoglycans decorin and biglycan, which are present in the extracellular matrix of cells, and transforming growth factor Beta-1 (TGF-B1), has been shown in many cancers. We investigated the significance of these molecules in NSCLC. Fasting serum levels of decorin, biglycan, and TGF-B1 were obtained from 48 newly diagnosed NSCLC patients and compared with those of 48 adult control subjects matched for age and demographics. Demographic data, baseline laboratory values, and ELISA results were compared between the groups. The median age was 65(39–83) similar in both groups. There was no relation between demographic and clinical parameters and the levels of decorin, biglycan, and TGF-B1 in the NSCLC group. However, in comparison to the control group, NSCLC patients had significantly higher levels of biglycan (42.55 ± 27.40 vs. 24.38 ± 12.05 ng/mL, p= 0.026) and TGF-B1 (15.55 ± 9.16 vs. 10.07 ± 7.8 pg/mL, p= 0.001), while decorin levels were significantly lower (6.64 ± 1.92 vs. 10.28 ± 3.13 ng/mL, p= 0.002). In the multivariate regression analysis; Decorin < 8.13 ng/mL (OR, 10.96; 95% CI: 3.440–34.958), current smoking (OR, 3.81; 95% CI: 1.320–10.998), COPD (OR, 43.6; 95% CI: 2.082–913.081), and lower BMI (OR, 1.22; 95% CI: 1.070–1.405, p= 0.003) were identified as independent predictive markers for NSCLC diagnosis. The decreased serum decorin level is an independent marker for NSCLC. Further studies are needed to investigate the prognostic significance of decorin on survival and its potential as a target in treatment. [ABSTRACT FROM AUTHOR]

Schistosomiasis mansoni leads to liver fibrosis that was believed to be irreversible once established leading to life-threatening complications in most cases. Praziquantel (PZQ) is considered the best drug in treatment, for decades. A universal approach towards drug repurposing is recommended, to be combined with other known drugs that may expand their efficiencies or/and discover new mechanisms of action of the used drugs. In this work Angiotensin-converting enzyme inhibitor (ACEI) "Lisinopril" was used alone or with Praziquantel (PZQ), versus long-duration PZQ (LD-PZQ) to test for their anti-fibrotic properties, applying Transforming Growth factor beta 1 (TGF-β1) as an indicator of activated fibrotic fibroblasts, to be automatically quantified within hepatic tissues of experimentally infected mice, using a software image analyzer. All treated groups showed significant reduction within TGF-β1 local hepatic expression, with the greatest reduction occurring among groups treated with LD-PZQ followed by those which received combined ACEI and PZQ. The highest TGF-β1 local hepatic expression values were obtained by groups receiving the classic anti-bilharzial single oral dose praziquantel (S.O.D PZQ) regimen, indicating its ineffectiveness as a monotherapeutic agent to minimize liver fibro-sclerotic threats in acute and chronic phases of schistosomiasis. Lisinopril alone or when combined with PZQ, succeeded in causing a drop in TGF-β1 hepatic expression as LD-PZQ regimen, thus increasing the chance of healing without hepatic scarring. Yet, its usage as an adjuvant to PZQ, instead of LD-PZQ has the additional benefit of not exposing the community to unwarranted resistance to PZQ, a drug that is to date still indispensable for the treatment of bilharziasis. [ABSTRACT FROM AUTHOR]

In the adult skeleton, the bone remodeling process involves a dynamic coordination between osteoblasts and osteoclasts, which is disrupted in diseases with high bone turnover rates and dysregulated transforming growth factor beta 1 (TGF-b1). However, little is known about how TGF-b1 signaling mediates bone resorption. Here, we described a pedigree with a heterozygous variant in TGF-b1 (R218C) that resulted in aberrant activation of TGF-b1 through an activating mechanism that caused Camurati-Engelmann disease (CED). We showed that CED patients have high levels of active Rho GTPases and the migration-related proteins Integrin b1 and Integrin b3 in their peripheral blood. HEK293T cells transfected with a plasmid encoding this mutant expressed high levels of TGF-b1 and active Rho GTPases. Furthermore, activation of Rho by TGF-b1 increased osteoclast formation and bone resorption, with increased migration of pre-osteoclasts, as well as cytoskeletal remodeling of pre-osteoclasts and mature osteoclasts. Importantly, pharmacological inhibition of Rho GTPases effectively rescued hyperactive TGF-b1-induced osteoclastogenesis in vitro. Overall, we propose that Rho GTPases mediate TGF-b1-induced osteoclastogenesis and suggest that Rho-TGF-b1 crosstalk is associated with high bone turnover in CED. [ABSTRACT FROM AUTHOR]

Despite its high prevalence, the cellular and molecular mechanisms of chronic obstructive pulmonary disease (COPD) are far from being understood. Here, we determine disease-related changes in cellular and molecular compositions within the alveolar space and peripheral blood of a cohort of COPD patients and controls. Myeloid cells were the largest cellular compartment in the alveolar space with invading monocytes and proliferating macrophages elevated in COPD. Modeling cell-to-cell communication, signaling pathway usage, and transcription factor binding predicts TGF-b1 to be a major upstream regulator of transcriptional changes in alveolar macrophages of COPD patients. Functionally, macrophages in COPD showed reduced antigen presentation capacity, accumulation of cholesteryl ester, reduced cellular chemotaxis, and mitochondrial dysfunction, reminiscent of impaired immune activation. [ABSTRACT FROM AUTHOR]

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive lung disease characterized by fibroproliferative matrix molecule accumulation, collagen deposition, and apoptosis. Activated leukocyte cell-adhesion molecule (ALCAM; CD166) is a celladhesion molecule that has been implicated in adhesive and migratory attribution, including leukocyte homing and trafficking and cancer metastasis. We investigated the role of ALCAM on pulmonary fibrosis development in murine models. Thus, a bleomycin-induced pulmonary fibrosis model was established with wild-type and ALCAM2/2 mice. Pulmonary fibrosis was also induced in transforming growth factor-b1 (TGF-b1)- transgenic mice that conditionally overexpress TGF-b1 upon doxycycline administration. In both models, observed reduced ALCAM levels in lung tissue and BAL fluid in pulmonary fibrosis-induced wild-type mice compared with control mice. We also observed reduced ALCAM expression in the lung tissue of patients with pulmonary fibrosis compared with normal lung tissue. ALCAM2/2 mice showed an exacerbated lung fibrosis response compared with wild-type mice, and this was accompanied by increased cell death. Further investigation for identification of the signaling pathway revealed that PI3K and ERK signaling pathways are involved in bleomycin-induced fibrosis. Collectively, these results highlight that ALCAM plays a protective role in the pathogenesis of pulmonary fibrosis that inhibits epithelial cell apoptosis through the PI3K-Akt signaling pathway. Our findings demonstrate the potential of ALCAM as a therapeutic target for IPF and may aid the development of new strategies for the management and treatment of patients with IPF. [ABSTRACT FROM AUTHOR]

Aim: The aim of this study was to evaluate the effects of standard culture medium and chondrogenic differentiation medium with PRP on chondrogenic differentiation of rabbit dental pulp-derived mesenchymal stem cells (rabbit DPSCs) that are transfected with transforming growth factor-beta 1 (TGF-B1) gene, based on the hypothesis of TGF- B1 and PRP can be effective on the chondrogenesis of stem cells. Materials and Methods: Rabbit DPSCs were characterized by using flow cytometry, immunofluorescent staining, quantitative Real Time Polymerase Chain Reaction (qRT-PCR) and differentiation tests. For the characterization, CD29, CD44 and CD45 mesenchymal cell markers were used. Rabbit DPSCs were transfected with TGF-B1 gene using electroporation technique in group 1; with PRP 10% in group 2; with chondrogenic medium in group 3; with both chondrogenic medium and PRP in group 4. DPSCs were cultured in medium with 10% inactive PRP in group 5, chondrogenic medium in group 6, chondrogenic medium with PRP 10% in group 7. SOX9, MMP13 and Aggrecan gene expression levels were evaluated in 3, 6, 12. and 24. days by qRT-PCR. Results: The expression levels of SOX9, MMP13 and Aggrecan were higher in group 2, 3 and group 7 in 3th day however in 24th day group 7 and group 2 were found higher. The expression levels changed by time-dependent. The extracellular matrix of the cells in experimental groups were positively stained with safranin O and toluidine blue. Conclusion: The combination in culture medium of TGF-B1 gene transfection and 10% PRP accelerates the chondrogenic differentiation of DPSCs. [ABSTRACT FROM AUTHOR]

Tetrahydrocurcumin (THC) is known to be the major metabolite of curcumin. The present study aimed to investigate the efficacy of THC and curcumin against hepatic fibrogenesis and the underlying responsible molecular mechanisms. Hepatic fibrogenesis was induced by intraperitoneal injection of CCl4 twice per week for 8 weeks. Daily oral administration of THC at 10 mg/kg and 50 mg/kg attenuated liver fibrogenesis more effectively than curcumin, as indicated by histopathological changes and a reduction in serum biochemical markers, collagen deposition, and fibrogenic molecules such as α-SMA and TGF-β1. THC exerted anti-fibrotic effects through the down-regulation of the TGF-β1/Smad and MAPKs signaling pathways. Moreover, IHC double-staining showed that THC diminished the autophagic process in activated HSCs in the liver. These findings suggest that THC alleviated hepatic fibrogenesis via inhibition of multiple signaling and autophagy suppression. THC could be used as a potent dietary supplement for the management of fibrotic liver diseases.

Esophageal cancer has an aggressive behavior with rapid tumor mass growth and frequently poor prognosis; it is known as one of the most fatal types of cancer worldwide. The identification of potential molecular markers that can predict the response to treatment and the prognosis of this cancer has been subject of a vast investigation in the recent years. Among several molecules, various angiogenic factors that are linked to the tumor development, growth, and invasion, such as VEGF, HGF, angiopoietin-2, IL-6, and TGF-B1, were investigated. In this paper, the authors sought to review the role of these angiogenic factors in prognosis and hypothesize how they can be used as a treatment target. [ABSTRACT FROM AUTHOR]

Hirsutella sinensis, cultured in vitro, is an attractive substitute for Cordyceps sinensis as health supplement. The aim of this study was to demonstrate whether H. sinensis mycelium (HSM) attenuates murine pulmonary fibrosis induced by bleo-mycin and to explore the underlying molecular mechanisms. Using lung fibrosis modle induced by intratracheal instillation of bleomycin (BLM; 4 mg/kg), we observed that the administration of HSM reduced HYP, TGF-b1 and the production of several pro-fibrosis cytokines (a-smooth muscle actin, fibronectin and vimentin) in fibrotic mice lung sections. Histopathological examination of lung tissues also demonstrated that HSM improved BLM-induced pathological damage. Concurrently, HSM supplementation markedly reduced the chemotaxis of alveolar macrophages and potently suppressed the expression of inflammatory cytokines. Also, HSM influenced Th1/Th2 and Th17/Treg imbalance and blocked the phosphorylation of mTOR pathway in vivo. Alveolar epithelial A549 cells acquired a mesenchymal phenotype and an increased expression of myofibroblast markers of differentiation (vimentin and fibronectin) after treatment with TGF-b1. HSM suppressed these markers and blocked the phosphorylation of mTOR pathway in vitro. The results provide evidence supporting the use of HSM in the intervention of pulmonary fibrosis and suggest that HSM is a potential therapeutic agent for lung fibrosis. [ABSTRACT FROM AUTHOR]

Tetrahydrocurcumin (THC) is known to be the major metabolite of curcumin. The present study aimed to investigate the efficacy of THC and curcumin against hepatic fibrogenesis and the underlying responsible molecular mechanisms. Hepatic fibrogenesis was induced by intraperitoneal injection of CCl 4 twice per week for 8 weeks. Daily oral administration of THC at 10 mg/kg and 50 mg/kg attenuated liver fibrogenesis more effectively than curcumin, as indicated by histopathological changes and a reduction in serum biochemical markers, collagen deposition, and fibrogenic molecules such as α-SMA and TGF-β1. THC exerted anti-fibrotic effects through the down-regulation of the TGF-β1/Smad and MAPKs signaling pathways. Moreover, IHC double-staining showed that THC diminished the autophagic process in activated HSCs in the liver. These findings suggest that THC alleviated hepatic fibrogenesis via inhibition of multiple signaling and autophagy suppression. THC could be used as a potent dietary supplement for the management of fibrotic liver diseases. [ABSTRACT FROM AUTHOR]

Dkk-3 is a member of the dickkopf protein family of secreted inhibitors of the Wnt pathway, which has been shown to enhance angiogenesis. The mechanism underlying this effect is currently unknown. Here, we used cultured HUVECs to study the involvement of the TGF-b and VEGF on the angiogenic effect of Dkk-3. Addition of hrDkk-3 peptide (1 or 10 ng/ml) to HUVECs for 6 or 12 h enhanced the intracellular and extracellular VEGF protein levels, as assessed by RTPCR, immunoblotting, immunocytochemistry and ELISA. The increase in the extracellular VEGF levels was associated to the VEGFR2 activation. Pharmacological blockade of VEGFR2 abrogated Dkk-3-induced endothelial cell tubes formation, indicating that VEGF is a molecular player of the angiogenic effects of Dkk-3. Moreover, Dkk-3 enhanced Smad1/5/8 phosphorylation and recruited Smad4 to the VEGF gene promoter, suggesting that Dkk-3 activated ALK1 receptor leading to a transcriptional activation of VEGF. This mechanism was instrumental to the increased VEGF expression and endothelial cell tubes formation mediated by Dkk-3, because both effects were abolished by siRNA-mediated ALK1 knockdown. In summary, we have found that Dkk-3 activates ALK1 to stimulate VEGF production and induce angiogenesis in HUVECs. [ABSTRACT FROM AUTHOR]

O presente estudo teve por objetivo avaliar a capacidade de concentração plaquetária e sua correlação com os níveis do fator de crescimento TGF-B1, a presença de leucócitos e de hemácias nos diferentes protocolos utilizados na obtenção do plasma rico em plaquetas (PRP) de equinos, através do método manual. Dez equinos, sadios, com idade média de 7 anos (±2,39), pesando em média 500kg (±67,1) foram utilizados neste estudo. Os protocolos testados variaram na velocidade e no tempo nas duas centrifugações. As variáveis analisadas nas amostras de PRP foram: concentração de plaquetas, presença de leucócitos e hemácias, e níveis de TGF-β1 quantificados pelo teste ELISA. Os protocolos testados não diferiram na capacidade de concentração de plaquetas e nos níveis de TGF-β1. Entretanto, houve diferença significava entre o protocolo I e os demais por este apresentar maior número de hemácias e leucócitos nas amostras de PRP, sendo por esse motivo considerado um protocolo inadequado para processamento do volume de sangue utilizado. Os demais protocolos podem ser utilizados para obtenção de PRP terapêutico em equinos.PRP is plasma that contains a high numbers of platelets and growth factors in a small volume. The aim of this study was to evaluate seven different protocols to obtain PRP by the manual method according to their capacity to concentrate platelets, leukocyte and erythrocyte contamination and correlation between platelet count and TGF-β1 growth factor levels in PRP samples. Ten healthy horses with a mean age of 7 years (±2.39), weighing on average 500 kg (±67.1) were used in this study. The protocols tested varied according to the speed and time used at the two centrifugations. PRP samples were analyzed regarding platelet concentration, leukocyte and erythrocyte contamination and TGF-β1 levels quantified by ELISA. No significant differences among protocols were observed regarding the ability to concentrate platelets and TGF-β1. However, protocol I showed significantly higher erythrocyte and leukocyte counts in PRP samples than the other protocols, reason why it was considered inadequate for the volume of blood processed in this experiment. The remaining protocols are suitable for extracting PRP.

Background : Oral submucous fibrosis (OSF) may be considered a collagen metabolic disorder resulting from areca-nut alkaloid exposure and individual variation in collagen metabolism. Due to the complexity of OSF pathogenesis, it is important to elucidate independent and interactive effects of polymorphisms of collagen-related genes on OSF risk. Materials and Methods : This study is focused on seven polymorphisms (SNPs) of transforming growth factor-beta-1 (TGF-beta-1) gene in patients with oral submucous fibrosis (OSF), belonging to south Indian ethnic extraction. The mean age at presentation was 43.9 years, range 23-72 years (n=50, M:F ratio, 2.6:1). DNA samples from 50 subjects of the same ethnic group and comparable demographic features who have had practiced the habit of areca-chewing of almost equal duration, but remained free of disease constituted the controls. All DNA samples were collected progressively and purified from peripheral blood employing standard protocols and tested for SNPs. They included two polymorphisms in the promoter region (C-509T and G-800A), three polymorphisms in exon-1 (Arg25Pro(G915C), Leu10Pro(T869C), Glu47Gly(A979G) and two in 5 ¢UTR regions (C→T(rs13306708) and G→A (rs9282871). The extracted DNA samples along with the primers underwent PCR amplification and the genotypic and allelic frequencies were calculated. All calculations were performed using the SPSS software. The PCR products were purified and subsequently sequenced using Flour S™ multi-imager system (Biorad). The sequenced data were analyzed using the BioEdit sequence analysis software. Results : Out of the seven polymorphisms analyzed, six such as two in the promoter region, three in exon-1 and one in 5¢UTR were found to have a " P" value above 0.05 and hence were not significant. The C→T transition (rs13306708) in the 5¢UTR region recorded a " P" value of 0.03 on comparison and hence was found to be significant. The allelic frequencies for this C→T transition in patients were 68.7% C and 31.2% T (27CC, 15CT, 8TT) and that in controls were 89.5% C and 10.4% T (42CC, 6CT, 2TT). Conclusions : The polymorphism in 5¢UTR C-T in TGF beta 1 gene has a significant association with OSF, being a prime determinant in the pro-angiogenic pathway which has got direct bearing with the pathophysiology of the disease. The proximity of this polymorphism to the transcription site and the associated risk involved is discussed.

Complement activation, an integral arm of innate immunity, may be the critical link to the pathogenesis of idiopathic pulmonary fibrosis (IPF). Whereas we have previously reported elevated anaphylatoxins--complement component 3a (C3a) and complement component 5a (C5a)--in IPF, which interact with TGF-β and augment epithelial injury in vitro, their role in IPF pathogenesis remains unclear. The objective of the current study is to determine the mechanistic role of the binding of C3a/C5a to their respective receptors (C3aR and C5aR) in the progression of lung fibrosis. In normal primary human fetal lung fibroblasts, C3a and C5a induces mesenchymal activation, matrix synthesis, and the expression of their respective receptors. We investigated the role of C3aR and C5aR in lung fibrosis by using bleomycin-injured mice with fibrotic lungs, elevated local C3a and C5a, and overexpression of their receptors via pharmacologic and RNA interference interventions. Histopathologic examination revealed an arrest in disease progression and attenuated lung collagen deposition (Masson's trichrome, hydroxyproline, collagen type I α 1 chain, and collagen type I α 2 chain). Pharmacologic or RNA interference-specific interventions suppressed complement activation (C3a and C5a) and soluble terminal complement complex formation (C5b-9) locally and active TGF-β1 systemically. C3aR/C5aR antagonists suppressed local mRNA expressions of tgfb2, tgfbr1/2, ltbp1/2, serpine1, tsp1, bmp1/4, pdgfbb, igf1, but restored the proteoglycan, dcn. Clinically, compared with pathologically normal human subjects, patients with IPF presented local induction of C5aR, local and systemic induction of soluble C5b-9, and amplified expression of C3aR/C5aR in lesions. The blockade of C3aR and C5aR arrested the progression of fibrosis by attenuating local complement activation and TGF-β/bone morphologic protein signaling as well as restoring decorin, which suggests a promising therapeutic strategy for patients with IPF. [ABSTRACT FROM AUTHOR]

Wprowadzenie: Czynniki wzrostowe IGF-1 oraz TGF-b1 są dobrze udokumentowanymi cytokinami regulującymi proliferencję komórek pęcherzykowych tarczycy w wielu doświadczeniach in vitro. Udowodniono dotychczas, że IGF-1 stymuluje mitogenezę komórkową tyreocytów, podczas gdy TGF-b1 ma właściwości hamujące proliferencję komórek pęcherzykowych tarczycy w warunkach doświadczalnych. Cel pracy: Celem badania była ocena zależności między stężeniem IGF-1 i TGF-b1 w surowicy krwi a wielkością tarczycy u dzieci z prawidłowym wydalaniem jodu z moczem. Materiał i metody: Badanie przeprowadzono w 2002 roku w 4 losowo wybranych szkołach podstawowych na terenie Białegostoku oraz w Dziecięcej Poradni Endokrynologicznej SP ZOZ Wojewódzkiego Szpitala Specjalistycznego w Białymstoku. Do udziału w badaniu zaproszono 480 dzieci w wieku 7-13 lat ze szkół oraz 120 pacjentów w tym samym przedziale wiekowym leczonych w Poradni Endokrynologicznej KJ i/lub tyroksyną przez minimum 12 miesięcy z powodu powiększenia tarczycy. U wszystkich dzieci przeprowadzono badanie przedmiotowe z oceną palpacyjną powiększenia tarczycy, wykonano badanie USG tarczycy oraz oceniono stężenie jodu w porannej zbiórce moczu metodą katalityczną Sanedell-Kolthoffa. W drugiej części badania na podstawie obliczonej wielkości tarczycy, opierając się na kryteriach WHO z 1997 roku dla powierzchni ciała i płci, badane dzieci podzielono na 2 podgrupy: z powiększoną tarczycą i z wielkością tarczycy w granicach normy. Do dalszych badań z wyodrębnionych podgrup zakwalifikowano dzieci w wieku 9-11 lat. W obu podgrupach dzieciom pobrano krew na oznaczenie stężenia TSH, IGF-1 i TGF-b1. Wyniki: Wartości średnie / mediana stężenia IGF-1 były istotnie statystycznie wyższe w grupie dzieci z powiększoną tarczycą w porównianiu do grupy dzieci z wielkością tarczycy w granicach normy (436,2 vs. 343,8 ng/ml, p=0,047). Wartości średnie / mediana TGF-b1 były istotnie statystycznie niższe w grupie dzieci z powiększoną tarczycą w porównaniu do grupy dzieci z wielkością tarczycy w granicach normy (17,8 vs. 23,9 ng/ml). Wnioski: Istotnie niższe stężenia TGF-b1 w surowicy krwi dzieci z powiększoną tarczycą w porównaniu do dzieci z prawidłową wielkością tarczycy oraz istnienie dodatniej korelacji stężeń IGF-1 z wielkością tarczycy (po wykluczeniu wpływu wieku i powierzchni ciała) wydają się potwierdzać istotną rolę IGF-1 i TGF-b1 w patomechanizmie powiększenia tarczycy.

Pulmonary fibrosis is a difficult to treat, often fatal disease whose pathogenesis involves dysregulated TGF-b1 signaling. CD4+CD25+FoxP3+ Regulatory T cells (Tregs) exert important effects on host tolerance and arise from naïve CD4+ lymphocytes in response to TGF-b1. However the precise contribution of Tregs to experimentally induced murine lung fibrosis remains unclear. We sought to better understand the role of Tregs in this context. Using a model of fibrosis caused by lung specific inducible overexpression of the bioactive form of the human TGF-b1 gene we find that Tregs accumulate in the lung parenchyma within five days of transgene activation and that this enhancement persists to at least fourteen days. Anti-CD25 Antibody mediated depletion of Tregs causes increased detection of soluble collagen and of intrapulmonary CD45+CD34+Col Ia1 fibrocytes. These findings are accompanied by enhanced local concentrations of the classical inflammatory mediators CD40L, TNF-a, and IL-1a, along with the neuroimmune molecule fibroblast growth factor 9 (FGF-9, also known as glial activating factor). FGF-9 expression localizes to structural cells and alveolar macrophages in this model and antibody mediated neutralization of FGF-9 results in attenuated detection of intrapulmonary collagen and fibrocytes without affecting Treg quantities. These data indicate that CD4+CD25+FoxP3+ Tregs attenuate TGF-b1 induced lung fibrosis and fibrocyte accumulation in part via suppression of FGF-9.

Objectives: This study aimed to investigate the effect of TGF-B1-transfected adipose-derived mesenchymal stem cell (AD-MSC) conditional medium (TGF-B1-CM) on CD44 expression and biological activities in MCF-7 and MDA-MB-231 cells., Methods: In the study, the experimental groups were created as a standard medium, AD-MSC-CM, TGF-B1-CM, and TGF-B1 recombinant protein. The medium and proteins specified in these groups were applied to MCF-7 and MDA-MB-231 cells separately at 24, 48 and 72 hours. Western blot and immunofluorescent staining were performed with antibodies suitable for CD44 and canonical smad signaling pathway analyses between groups. Cellular proliferation in MCF-7 and MDA-MB-231 cells was measured by MTT. Biological activity analyses such as apoptosis, cell cycle, proliferation, DNA damage, and membrane depolarization between groups were tested on the Muse Cell Analyzer using appropriate kits. Cellular migration between groups was determined by showing cells that migrated to the scar area with in vitro scar formation. Statistics were performed with GraphPad Prism 8.02 software., Results: It was determined that TGF-B1-CM activates the smad signaling pathway in MCF-7 and MDA-MB-231 cells. TGF-B1-CM increased pSMAD2/3 expression and decreased SMAD4 expression in breast cancer cells. A decrease in CD44 expression was found at points of increase in pSMAD2/3 expression. Decreased expression of SMAD4 in breast cancer cells with TGF-B1-CM was associated with decreased expression of CD44. In MCF-7 and MDA-MB-231 cells, TGF-B1-CM was found to increase apoptosis, decrease proliferation, disrupt membrane depolarization, and arrest cells at G0/G1 stage. TGF-B1-CM suppressed MCF-7 and MDA-MB-231 migrations., Conclusion: SMAD4-targeted therapeutic strategies may be considered to suppress CD44 expression in breast cancer cells. Both the anti-tumorigenic factors released by AD-MSCs and the secretomes obtained as a result of supporting these factors with the overexpression of TGF-B1, severely suppressed breast cancer cells. With this study, it was planned to obtain a targeted biological product that suppresses breast cancer cells in vitro ., Competing Interests: None., (AJSC Copyright © 2022.)

ABSTRACT Macrophages are present in large numbers and display specific and distinct phenotypes during the various phases of tissue repair. However, their role following tendon injury and during repair has never been investigated. We injected C57BL/6 mice daily for 4 days with liposome-encapsulated clodronate to deplete circulating monocytes/macrophages. Placebo mice were injected with PBS. The left Achilles tendons of the mice were transversely sectioned and sutured using the 8-strand technique. Macrophage accumulation and cell proliferation were significantly lower in the tendons of clodronate-treated mice than in those of PBS-treated mice on days 3 and 7 post-injury. TGF-β1 staining was significantly more intense in the tendons of PBS-treated mice on day 7 post-injury. Edema and the dry mass of the Achilles tendons were also higher in the PBS-treated mice on days 7 and 14 post-injury. No differences in absolute strength and stiffness were observed, but Young's modulus and maximal stress were significantly greater for tendons from the clodronate-treated mice than those from PBS-treated mice after 14 days of tendon repair. Overall, our findings showed that macrophages promote cell proliferation and extracellular matrix accumulation but their presence leads to inferior ultimate tensile strength of the Achilles tendons. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:279-285, 2014. [ABSTRACT FROM AUTHOR]

PRP is plasma that contains a high numbers of platelets and growth factors in a small volume. The aim of this study was to evaluate seven different protocols to obtain PRP by the manual method according to their capacity to concentrate platelets, leukocyte and erythrocyte contamination and correlation between platelet count and TGF-β1 growth factor levels in PRP samples. Ten healthy horses with a mean age of 7 years (±2.39), weighing on average 500 kg (±67.1) were used in this study. The protocols tested varied according to the speed and time used at the two centrifugations. PRP samples were analyzed regarding platelet concentration, leukocyte and erythrocyte contamination and TGF-β1 levels quantified by ELISA. No significant differences among protocols were observed regarding the ability to concentrate platelets and TGF-β1 . However, protocol I showed significantly higher erythrocyte and leukocyte counts in PRP samples than the other protocols, reason why it was considered inadequate for the volume of blood processed in this experiment. The remaining protocols are suitable for extracting PRP. [ABSTRACT FROM AUTHOR]

Recent data show that regulatory cells with transforming growth factor (TGF)-β1-dependent activity are able to restore self-tolerance in overtly diabetic non-obese diabetic (NOD) mice. Thus, TGF-β1 seems to have a relevant role in protection from autoimmune diabetes. Our aim was to investigate the possible significance of serum TGF-β1 measurement in the natural history of diabetes in NOD mice, as well as in children positive for at least one islet-related antibody. Serum TGF-β1 (both total and active) was measured by enzyme-linked immunosorbent assay at monthly intervals in 26 NOD mice during the spontaneous development of diabetes and, on a yearly basis, in nine siblings of patients with type 1 diabetes (T1D) with a follow-up of 4 years. Diabetes appeared between the 12th week of age and the end of the study period (36 weeks) in 17 mice. TGF-β1 serum level variations occurred in the prediabetic period in both NOD mice and humans and diabetes diagnosis followed a continuing reduction of active TGF-β1 (aTGF-β1) serum levels. In mice, aTGF-β1 serum levels measured at 4 weeks of age correlated positively with severity of insulitis, and negatively with percentage of insulin-positive cells. Our findings suggest that in NOD mice serum TGF-β1 levels during the natural history of the diabetes reflect the course of islet inflammation. The measurement of aTGF-β1 in islet-related antibody-positive subjects may provide insights into the natural history of prediabetic phase of T1D. [ABSTRACT FROM AUTHOR]

The aim of this study was designed to investigate the possible beneficial effects of the angiotensin (ang) II T1 (AT1) receptor blocker, irbesartan (Irb), and the alpha lipoic acid (ALA) in streptozotocin (STZ)-induced diabetic nephropathy (DNP) in rats. The rats were randomly allotted into one of five experimental groups: A, control; B, diabetic untreated; C, diabetic treated with Irb; D, diabetic treated with ALA; and E, diabetic treated with Irb + ALA; each group contains 10 animals. B, C, D, and E groups received STZ. Diabetes was induced in four groups by a single intraperitoneal injection of STZ (50 mg/kg, freshly dissolved in 5 mmol/L citrate buffer, pH 4.5). The rats in Irb-, ALA-, and Irb + ALA-treated groups were given Irb (5 mg/kg), ALA (in a dose of 3 mg/kg), and Irb + ALA (in a dose of 2.5 + 1.5 mg/kg) once a day orally by using intragastric intubation for 12 weeks starting 2 days after STZ injection, respectively. Treatment with ALA and especially Irb reduced the glomerular size; thickening of capsular, glomerular, and tubular basement membranes; increased amounts of mesangial matrix and tubular dilatation as compared with diabetic-untreated rats. Notably, the better effects were obtained when Irb and ALA were given together. We conclude that Irb, ALA, and especially Irb + ALA therapy causes renal morphologic improvement after STZ-induced diabetes in rats. We believe that further preclinical research into the utility of Irb and ALA treatment, alone or its combination, may indicate its usefulness as a potential treatment in DNP. [ABSTRACT FROM AUTHOR]

Morphine is a commonly prescribed analgesic for wound pain. Previous studies have shown that morphine enhances accumulation of collagen in cultured fibroblasts. Because fibroblasts are important for the remodeling of connective tissue in incisional wound, this study investigates the biological effects of morphine on cutaneous collagen content and wound tensile strength. A full-thickness incisional wound (2 cm in length) was created on the dorsum of mice followed by treatment with placebo or morphine (5 and 20 mg/kg/day, i.p.). Fourteen days later, tensile strength of the healed incisional wound was measured using a tensiometer. Protein expression of transforming growth factor (TGF)-β1 and matrix metalloproteinases (MMP)-2 in the incisional wound tissue was analyzed. Degree of tissue remodeling and levels of collagen were determined by histological examination and a dye-binding collagen assay, respectively. Morphine enhanced the breaking strength of incisional wound 14 days after treatment (92 ± 10, 102 ± 10 and 134 ± 12 mg for control, morphine 5 mg/kg/day and morphine 20 mg/kg/day, respectively; P = 0.03, n = 6–7). Protein expression of TGF-β1 and MMP-2 was significantly enhanced in mice treated with morphine. Histological examination of the wound tissue showed evidence of increased thickness of the cutaneous fibrous layer and deposition of collagen in the high-dose morphine treatment group. Collagen assays also demonstrated that tissue concentrations of collagen were significantly increased in the wound tissue of morphine-treated animals on day 2 of drug treatment. The present study demonstrates that systemic administration of morphine enhances tissue collagen deposition in the cutaneous tissue, thereby increasing the tensile strength of the incisional wound. [ABSTRACT FROM AUTHOR]

Synovium is considered a candidate source of cells for cartilage tissue engineering. Compared with mesenchymal stem cells (MSCs) from other sources, synovium-derived stem cells (SDSCs) have a higher capacity for chondrogenic differentiation. Our objective was to define cocktails of growth factors that support the growth and chondrogenic differentiation of SDSCs in chemically defined medium. We established a fast and highly selective technique of negative isolation of SDSC populations. The individual and combined effects of three growth factors—transforming growth factor-β1 (TGF-β1), insulin-like growth factor I (IGF-I), and basic fibroblast growth factor (FGF-2)—were evaluated in serum-free pellet cultures of SDSCs for the chondrogenesis of SDSCs using histology, biochemical analysis, and real-time RT-PCR. In vitro studies identified TGF-β1 as the key factor for both the growth and chondrogenesis of SDSCs. The highest rates of SDSC growth were observed with the synergistic interaction of all three factors. With respect to chondrogenic differentiation of SDSCs, the interaction of TGF-β1 and IGF-I applied simultaneously was superior to the sequential application of these two factors or any other combination of growth factors studied. Based on these findings, we propose a two-step protocol for the derivation of chondrogenic SDSCs: a cocktail of TGF-β1, IGF-I, and FGF-2 is applied first to induce cell growth followed by a cocktail of TGF-β1 and IGF-I applied to induce chondrogenesis. [ABSTRACT FROM AUTHOR]

Platelet rich plasma (PRP) is being used with increased frequency in many surgical procedures for its known benefits of accelerated surgical wound site healing. Speculations in its efficacy in the presence of anti-platelet therapy have been proposed. To aid in defining a quality platelet rich plasma product in the presence of acetylsalicylic acid (ASA) and Plavix (clopidogrel bisulfate), we investigated three (3) groups (n=18) of cardiac surgical patients receiving PRP. Platelet function test, platelet concentration, and quantification of growth factors (PDGF-bb and TGF-b1) were evaluated. Results showed no statistical evidence of decreased growth factors delivered to the surgical wound site in the presence of acetylsalicylic acid (ASA) and/or Plavix (clopidogrel bisulfate). Evidence in this pilot study supports the use of PRP for patients receiving Plavix and aspirin therapy without compromising the quantity of specific growth factors delivered to a wound site. [ABSTRACT FROM AUTHOR]

Although previous studies have demonstrated that diabetic nephropathy is attributable to early extracellular matrix accumulation in glomerular mesangial cells, the molecular mechanism by which high glucose induces matrix protein deposition remains not fully elucidated. Rat mesangial cells pretreated with or without inhibitors were cultured in high-glucose or advanced glycation end product (AGE) conditions. Streptozotocin-induced diabetic rats were given superoxide dismutase (SOD)-conjugated propylene glycol to scavenge superoxide. Transforming growth factor (TGF)-β1, fibronectin expression, Ras, ERK, p38, and c-Jun activation of glomerular mesangial cells or urinary albumin secretion were assessed. Superoxide, not nitric oxide or hydrogen peroxide, mediated high glucose- and AGE-induced TGF-β1 and fibronectin expression. Pretreatment with diphenyliodonium, not allopurinol or rotenone, reduced high-glucose and AGE augmentation of superoxide synthesis and fibronection expression. High glucose and AGEs rapidly enhanced Ras activation and progressively increased cytosolic ERK and nuclear c-Jun activation. Inhibiting Ras by manumycin A reduced the stimulatory effects of high glucose and AGEs on superoxide and fibronectin expression. SOD or PD98059 pretreatment reduced high-glucose and AGE promotion of ERK and c-Jun activation. Exogenous SOD treatment in diabetic rats significantly attenuated diabetes induction of superoxide, urinary albumin excretion, 8-hydroxy-2′-deoxyguanosine, TGF-β1, and fibronectin immunoreactivities in renal glomerular mesangial cells. Ras induction of superoxide activated ERK-dependent fibrosis-stimulatory factor and extracellular matrix gene transcription of mesangial cells. Reduction of oxidative stress by scavenging superoxide may provide an alternative strategy for controlling diabetes-induced early renal injury.Kidney International (2006) 69, 1593–1600. doi:10.1038/sj.ki.5000329; published online 29 March 2006 [ABSTRACT FROM AUTHOR]

Atopic dermatitis (AD) is a chronic inflammatory skin disease with immunopathologic features that vary depending on the duration of the lesion. The dermis of lesional skin of AD patients shows an increased number of inflammatory cells such as mast cells, eosinophils and mononuclear cells and superficialStaphylococcus aureuscolonization.The purpose of this study was to determine the effects of peptidoglycan (PEG) fromS. aureuson mast cell induction in murine skin.PEG was applied to barrier-disrupted abdominal skin of mice every 5 days and the number of mast cells in the abdominal skin was counted 20 days after the first application. The cytokine response was investigated by RT-PCR and immunohistologic analysis.The number of mast cells in the skin of mice treated with PEG was increased significantly compared with that of mice given phosphate-buffered saline. In addition, application of PEG to the abdominal skin increased the expression of mRNA for transforming growth factor-β1 (TGF-β1), which supports mast cell migration, but not that for IL-3 or stem cell factor, which support both mast cell proliferation and mast cell migration. Immunohistologic analysis demonstrated that levels of TGF-β1 transcripts corresponded with those of protein synthesis in the epidermis. TGF-β1 was found to be highly expressed in keratinocytes of the basal epidermis of PEG-treated skin. Furthermore, intraperitoneal injection of anti-TGF-β1 antibodies neutralized the induction of mast cells into the skin.These results suggest that PEG may have the ability to induce an increase in mast cell numbers in the skin through TGF-β1 production by epidermal keratinocytes. Skin inflammation might therefore be linked to colonization withS. aureusin AD patients. [ABSTRACT FROM AUTHOR]

Postoperative infections after hepatectomy sometimes lead to fatal hepatic failure, but the mechanism of the hepatic failure is unclear. Wistar rats underwent 90% hepatectomy, and were then divided into three groups: (i) the SAL group, injected with normal saline; (ii) the LPS group, injected with lipopolysaccharide (LPS) every day for 1 week; and (iii) the LPS plus TGF-Ab (LPS+TGF-Ab) group, injected with LPS with anti-transforming growth factor-β1 (TGF-β1) antibody. We investigated survival rates, TGF-β1 expression in the liver, liver regeneration by proliferating cell nuclear antigen labeling index, hepatocyte apoptosis by single stranded DNA labeling index, and perisinusoidal fibrosis using Masson's trichrome staining. The LPS group (30.4%) had a significantly lower survival rate than the SAL group (84%) and tended to be lower than the LPS+TGF-Ab group (49.4%). Liver regeneration in the LPS group was significantly lower than in the other groups. In the LPS group, hepatocyte apoptosis and perisinusoidal fibrosis was significantly more remarkable, and TGF-β1 expression was significantly higher than in the SAL group. TGF-β1 enhanced by LPS plays an important role in the mechanism of hepatic failure by infections after hepatectomy, especially in inhibition of liver regeneration, and induction of hepatocyte apoptosis and perisinusoidal fibrosis. [ABSTRACT FROM AUTHOR]

Estradiol reverses renal injury in Alb/TGF-β1 transgenic mice.Background. Men with chronic renal disease progress more rapidly to renal failure than do women. Tranforming growth factor-β (TGF-β) plays a central role in promoting progressive renal injury, in part due to transcriptional effects mediated by cooperation between Smad proteins and the transcription factor Sp1. Estrogen negatively regulates Sp1 activity and reverses the stimulatory effects of TGF-β on type IV collagen synthesis and cellular apoptosis in cultured mesangial cells. We hypothesized that the ability of estradiol to reverse the effects of TGF-β underlies gender dimorphism in the progression of chronic renal disease.Methods. We studied Alb/TGF-β transgenic mice, which overexpress TGF-β1 and develop proteinuria and progressive glomerulosclerosis. We implanted a sustained-release estradiol pellet or a placebo pellet into control and Alb/TGF-β transgenic mice at 2 weeks of age. Animals were sacrificed at 5 weeks, at which time urine, blood, and renal tissue were obtained for study.Results. The sustained-release estradiol pellet achieved a physiologic concentration of estradiol. TGF-β levels were higher in estradiol-treated mice compared to placebo-treated mice. Proteinuria was reduced in estradiol-treated Alb/TGF-β mice compared to placebo-treated transgenic mice. Mesangial expansion and closure of capillary loops with enhanced glomerular deposition of type I collagen, type IV collagen, and tissue inhibitor of metalloproteinase (TIMP-2) was observed in glomeruli of placebo-treated transgenic mice. Estrogen therapy reversed these abnormalities.Conclusion. Administration of estradiol to Alb/TGF-β transgenic mice, which overexpress TGF-β, ameliorated progressive renal injury. The ability of estradiol to reverse the pro-fibrotic effects of TGF-β, both in vitro and in vivo, may underlie the sexual dimorphism in renal disease progression observed in humans. [ABSTRACT FROM AUTHOR]

We have examined the effect of transforming growth factor β1 (TGF-β1) and overexpression of the Smad4 gene on the phenotype of Car C, a ras mutated highly malignant spindle carcinoma cell line. TGF-β1-treated Car C cells overexpressing Smad4 spread with a flattened morphology with membrane ruffles abundant in vinculin and show a reduction in their invasive abilities. TGF-β1 treatment and overexpression of Smad4 also enhanced the production of PAI-1 measured by the activation of the p3TP-lux reporter gene containing a PAI-1-related promoter. This activation was abolished with a dominant-negative Smad4 construct. These results lead us to conclude that both TGF-β1 and Smad4 overexpression reduce the invasive potential of Car C cells, probably via the Smad pathway. [Copyright &y& Elsevier]